%0 Journal Article
%T Fingerprinting of Natural Product by Eastern Blotting Using Monoclonal Antibodies
%A Hiroyuki Tanaka
%A Waraporn Putalun
%A Yukihiro Shoyama
%J Chromatography Research International
%D 2012
%I Hindawi Publishing Corporation
%R 10.1155/2012/130732
%X We succeeded in developing the fingerprint of natural product by eastern blotting using monoclonal antibodies. After developing and separating them on a TLC plate, solasodine glycosides are oxidized by NaIO4 and reacted with a protein to give conjugates which are recognized with anti-solamargine monoclonal antibody (MAb). Anti-solamargine MAb having wide cross-reactivity can stain and detect all solasodine glycosides by fingerprint. Different sensitivity between solamargine and solasonine was observed. The detection limit was 1.6£¿ng of solasonine. The hydrolysed products of solamargine were determined by fingerprint of eastern blotting compared to their Rf values depending on the sugar number. Fingerprint by eastern blotting using anti-ginsenoside Rb1 MAb distinguished the formula containing ginseng prescribed in traditional Chinese medicine. By double-staining of ginsenosides it is possible to suggest that the staining color shows the pharmacological activity, such as the purple bands indicate ginsenosides having stimulation activity, and the blue color indicated compound like ginsenosides possessed the depression affect for the central nervous system (CNS), respectively. 1. Introduction In the recent rapid development of the molecular biosciences and their biotechnological applications, immunoassay systems using monoclonal antibody (MAb) against drugs and small molecular weight bioactive compounds have become an important tool for studies on receptor binding analysis, enzyme assay, and quantitative and/or qualitative analytical techniques in animals or plants; owing to their specific affinity. Previously we prepared various kinds of MAb against natural products like forskolin [1], solamargine [2], crocin [3], marihuana compound [4], opium alkaloids [5], ginsenosides [6, 7], berberine [8], sennosides [9], paeoniflorin [10], glycyrrhizin [11, 12], ginkgolic acid [13], aconitine alkaloid [14], baicalin [15], and so on, and developed individual competitive enzyme-linked immunosorbent assay (ELISA) as a high sensitive, specific, and simple methodology. Western blotting is widely used as an immunostaining technique to detect high-molecular compounds like peptides and proteins based upon an antigen-antibody reaction. However, low-weight molecular compounds had not been previously analyzed by western blotting. We succeeded the blotted staining of solasodine glycoside on PVDF membrane using MAb after developed and separated solasodine glycosides on TLC and called as new-western blotting [16]. From this evidence, we applied this new methodology to licorice
%U http://www.hindawi.com/journals/cri/2012/130732/